中文摘要：

目的：建立培养成年小鼠心肌细胞模型，给予高糖高脂刺激观察是否可以引起心肌细胞损伤和凋亡，并分析其机制。 方法：分离培养成年小鼠心肌细胞，以最小必需培养基（minimum essential medium，MEM）加20mmol／L甘露醇作为正常对照培养基（正常对照组，n=10）；以MEM＋高糖（20mmol／L葡萄糖）高脂（50μmol／L软脂酸）培养模拟糖尿病（高糖高脂组，n=10），心肌细胞分别在两种培养基中培养24h后测定指标。 结果：高糖高脂组乳酸脱氢酶（LDH）、细胞凋亡蛋白酶caspase-3活性显著升高，过氧化亚硝基损伤标志物硝基酪氨酸生成增多，同时心肌诱生型一氧化氮合酶（iNOS）蛋白表达水平和一氧化氮（NO）生成增多，烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶蛋白表达水平和超氧阴离子生成增多，与正常对照组比较，差异均有统计学意义（P均〈0．01）。 结论：在培养的成年小鼠心肌细胞，高糖高脂可引起心肌细胞损伤和凋亡，这主要是由于高糖高脂引起的iNOS大量表达所生成的NO和NADPH氧化酶表达增高生成的超氧阴离子反应产生的大量过氧化亚硝基所诱发。

英文摘要：

Objective：To observe the oxidative stress injury and apoptosis in adult mouse cardiomyocytes induced by high glucose and high fatty acid culture, and to investigate its mechanism. Methods：Cardiomyocytes were isolated from adult mouse. The cell culture was divided into two groups. Control group：MEM with 20 mM mannitol, and High glucose and high fatty acid group： MEM with 20 mM glucose and 50 uM palmi-tare. Cardiomyocytes were cultured in those two kinds of medium for 24 hours and then the cardiomyocytes and conditioned medium were collected respectively for different assays. Results： Compared with Control group, High glucose and high fatty acid group showed increased activity of LDH which indi-cated the injury of cardiomyocytes, and increased activity of caspasc-3 which implied for cardiomyocytes apoptosis. High glucose and high fatty acid group also showed increased expression of iNOS and nitric oxide production, NADPH oxidase and superoxide anion production. Those substances could increase the damage of eardiomyocyte. Conclusion：High glucose and high fatty acid culture could up-regulate the expression of iNOS and NADPH thus increasing NO and superoxide anion production. Those substances could generate peroxynitrite and therefore induce the injury and apoptosis in cultured adult mouse cardiomyocytes.